Isolation of Hem3 mutants from Candida albicans by sequential gene disruption View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1989-05

AUTHORS

Myra B. Kurtz, Jean Marrinan

ABSTRACT

Molecular methods for directed mutagenesis in Candida albicans have relied on a combination of gene disruption by transformation to inactivate one allele and UV-induced mitotic recombination or point mutation to produce lesions in the second allele. An alternate method which uses two sequential gene disruptions was developed and used to construct a C. albicans mutant defective in a gene essential for synthesizing tetrapyrrole (uroporphyrinogen I synthase). The Candida gene was cloned from a random library by complementation of the hem3 mutation in Saccharomyces cerevisiae. The complementing region was limited to a approximately 2.0 kb fragment by subcloning and a Bg/II site was determined to be within an essential region. Linear fragments containing either the Candida URA3 or LEU2 gene inserted into the Bg/II site were used to disrupt both alleles of a leu2, ura3 mutant by sequential transformation. Ura+, Leu+ heme-requiring strains were recovered and identified as hem3 mutants by Southern hybridization, transformation to heme independence by the cloned gene, and enzyme assays. More... »

PAGES

47-52

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf00330941

DOI

http://dx.doi.org/10.1007/bf00330941

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1024596262

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/2671651


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